1. Identification of N 6 -methyladenosine reader proteins.
- Author
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Zhou KI, Liu N, and Pan T
- Subjects
- Adenosine analysis, Adenosine genetics, Adenosine metabolism, Binding Sites physiology, Electrophoresis, Polyacrylamide Gel methods, HEK293 Cells, Humans, Protein Binding physiology, RNA-Binding Proteins analysis, Adenosine analogs & derivatives, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism
- Abstract
The reversible N
6 -methyladenosine (m6 A) modification of eukaryotic messenger RNAs (mRNAs) is a widespread regulatory mechanism that impacts every step in the mRNA life cycle. The effect of m6 A on mRNA fate depends on the binding of "m6 A reader" proteins - RNA binding proteins that specifically bind to RNAs containing m6 A. Here, we describe an RNA pull-down method that can be used to identify novel m6 A reader proteins starting from a known m6 A-modified site in cellular or viral RNA. We further describe how a combination of immunoprecipitation-based sequencing methods can be used to identify m6 A-modified sites bound by an m6 A reader protein on a transcriptome-wide level. The discovery of new m6 A reader proteins and their m6 A-modified targets would provide further insight into the mechanisms and functions of m6 A in the cell., (Copyright © 2017 Elsevier Inc. All rights reserved.)- Published
- 2017
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